| Products/Services Used | Details | Operation |
|---|---|---|
| Gene Synthesis | Anti-DYX1C1 antibody (CT299) was raised against maltose-binding protein fused to the 206-residue DYX1C1 sequence predicted by JGIChlamydomonas genome version 4 (http:// genome.jgi.doe.gov/Chlre4/Chlre4.home.html). This DYX1C1 antigen was synthesized by GenScript USA Inc. and corresponds to residues 1–142 and 324–350 of both the DYX1C1 sequence determined in this study and the full-length protein sequence predicted by PhytozomeChlamydomonas genome version 5.5 (https://phytozome.jgi.doe.gov/pz/portal.html#! info?alias=Org_Creinhardtii). Phenotypic rescue ofpf23using wild-type DYX1C1 cDNA and the pGenD vector was carried out as described previously [18, 72]. The wild-type DYX1C1cDNA was synthesized by GenScript Japan (http://www.genscript.jp/gene_synthesis.html), and inserted into theNdeIEcoRI sites of the modified pGenD vector, which contains theAPHVIIIgene conferring paromomycin resistance toChlamydomonas. | Get A Quote |
Cytoplasmic assembly of ciliary dyneins, a process known as preassembly, requires numerous non-dynein proteins, but the identities and functions of these proteins are not fully elucidated. Here, we show that the classical Chlamydomonas motility mutant pf23 is defective in the Chlamydomonas homolog of DYX1C1. The pf23 mutant has a 494 bp deletion in the DYX1C1 gene and expresses a shorter DYX1C1 protein in the cytoplasm. Structural analyses, using cryo-ET, reveal that pf23 axonemes lack most of the inner dynein arms. Spectral counting confirms that DYX1C1 is essential for the assembly of the majority of ciliary inner dynein arms (IDA) as well as a fraction of the outer dynein arms (ODA). A C-terminal truncation ... More
